本文關(guān)鍵詞： 玉米 秸稈降解 低溫 復(fù)合菌系 菌種組成 穩(wěn)定性　出處：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2016年博士論文　論文類型：學(xué)位論文

【摘要】：我國玉米秸稈資源非常豐富,但目前對(duì)其利用很不充分。秸稈還田可以提高土壤養(yǎng)分,實(shí)現(xiàn)農(nóng)業(yè)生產(chǎn)的可持續(xù)發(fā)展。但在我國北方春玉米區(qū)秋收后低溫等氣候因素限制還田秸稈的當(dāng)季腐解,秸稈在土壤中腐解轉(zhuǎn)化的時(shí)間較長(zhǎng),不僅不能作為當(dāng)季作物的肥源還會(huì)影響后季作物播種質(zhì)量。為加快原位還田玉米秸稈的分解速率,提高腐解質(zhì)量,本研究從低溫(年平均氣溫-7℃-8℃)生態(tài)環(huán)境中采集長(zhǎng)年秸稈還田土壤、牛羊糞、腐爛樹葉等樣品為菌源材料,通過富集培養(yǎng)、碳源限制性繼代培養(yǎng)與低溫馴化等過程,篩選低溫條件(4℃～10℃)下高效降解玉米秸稈的復(fù)合菌系,并明確其秸稈分解特性、菌種組成特性、穩(wěn)定性和適應(yīng)性以及應(yīng)用效果,為北方地區(qū)低溫條件(4℃～10℃)下玉米秸稈快速、高效腐解以及合理利用提供理論依據(jù),并提供菌株資源。主要研究結(jié)果如下：1.玉米秸稈低溫高效降解復(fù)合菌系的篩選：利用富集培養(yǎng)、限制性繼代培養(yǎng)、低溫逐代馴化等技術(shù)對(duì)128份采集于高寒地區(qū)玉米秸稈還田土壤、牛羊糞、腐爛樹葉等菌源材料進(jìn)行篩選,獲得7個(gè)低溫(10℃)、中溫(15℃)雙高效型菌系和20個(gè)低溫(10℃)高效型菌系。進(jìn)一步通過對(duì)不同代數(shù)菌系玉米秸稈降解率及纖維素酶活性動(dòng)態(tài)分析,最終獲得一組可在低溫條件(4℃～10℃)下高效產(chǎn)纖維素酶,并穩(wěn)定降解玉米秸稈的復(fù)合菌系GF-20,其在接種后12h內(nèi)進(jìn)入對(duì)數(shù)生長(zhǎng)期,分泌纖維素酶,分解玉米秸稈成簡(jiǎn)單酯類或烴類。在10℃低溫條件下培養(yǎng)15d,玉米秸稈降解率達(dá)到32.29%,較對(duì)照復(fù)合菌系No.8(本研究小組2013年篩選,溫度15℃條件下降解率為22.02%)高出10.27%。2.玉米秸稈低溫高效降解復(fù)合菌系菌種組成多樣性：復(fù)合菌系GF-20由22種細(xì)菌和4種真菌組成。細(xì)菌組成分別為Azonexus hydrophilus、Azospira oryzae、Variovorax boronicumulans、Hydrogenophaga caeni、Herbaspirllum frisingense、Arcobacter cloacae、 Pseudomonas fuscovaginae、Cellvibrio mixtus subsp.mixtus、Seleniivibrio woodruffii、 Rhodococcus qingshengii、Terribacillus saccarophilus、Planococcus antarcticus、Bacillus licheniformis、Bacillus tequilensis、Clostridium populeti、Clostridium xylanolyticum、 Paludibacter propionicigenes、Pedobacter agri、Sphingobacterium cladoniae和Uncultured bacterium等。真菌主要為L(zhǎng)otharella globosa、Trichosporon sp.、Trichosporon loubieri和Chlamydomonas leiostraca。3.玉米秸稈低溫高效降解復(fù)合菌系穩(wěn)定性及適應(yīng)性：復(fù)合菌系GF-20在分解玉米秸稈過程中區(qū)系變化較穩(wěn)定,關(guān)鍵菌株穩(wěn)定存在；在不同溫度(4℃-30℃)和pH值(6.0-9.0)條件下繼代培養(yǎng)、在變溫條件(0℃-40℃)下連續(xù)培養(yǎng),復(fù)合菌系性質(zhì)、功能與組成均保持穩(wěn)定,具有較強(qiáng)的穩(wěn)定性和適應(yīng)性；碳源為玉米秸稈、氮源為硫酸銨時(shí)復(fù)合菌系GF-20菌種組成更穩(wěn)定,誘導(dǎo)復(fù)合菌系產(chǎn)纖維素酶,促進(jìn)底物分解；復(fù)合菌系GF-20的臨界稀釋梯度為1014倍。4.玉米秸稈低溫高效降解復(fù)合菌系應(yīng)用：在接種量10%、pH值6.5、玉米秸稈和硫酸銨添加比40：1、溫度10℃、滲透壓濃度2.0%條件下發(fā)酵制成的腐解菌劑GF-20具有良好的促分解效果。在4℃、10℃和30℃條件下腐解60d,玉米秸稈降解率分別為29.00%、36.75%和59.47%,較對(duì)照綠康菌劑高出7.21%、9.78%和10.26%；堿解氮、有效磷、速效鉀和有機(jī)質(zhì)含量分別增加0%-5.32%、5.26%-14.17%、2.07%-3.57%和0.44%～2.94%；顯著提高土壤過氧化氫酶、脲酶、堿性磷酸酶、蔗糖酶、B-葡萄糖苷酶和纖維素酶活性；增加土壤微生物豐富度指數(shù)與多樣性指數(shù),新增Candidatus saccharibacteria和Planctomycetia等微生物類群,促進(jìn)Herbaspirillum lusitanum、Bacillus sp.、Cellvibrio fibrivorans和Melanocarpus albomyces等功能微生物數(shù)量的增加,且菌劑GF-20對(duì)玉米種子發(fā)芽率無影響。玉米秸稈低溫高效降解復(fù)合菌系GF-20具有穩(wěn)定的玉米秸稈分解活性和菌種組成穩(wěn)定性,能在低溫條件(4℃～10℃)下有效加快秸稈分解進(jìn)程,提高土壤速效養(yǎng)分含量及酶活性,增加土壤微生物群落多樣性,具有良好的開發(fā)潛力和應(yīng)用前景。
[Abstract]:Corn straw resources are very rich in China, but its use is not sufficient. Returning straw can increase soil nutrients, to achieve the sustainable development of agricultural production. But in the area of northern China spring maize harvest after low temperature climatic factors such as seasonal restrictions of straw decomposition, stalk rot in soil solution into a long time not only can not be used as fertilizer, when will affect crop after crop sowing quality. In order to speed up the decomposition rate in situ returning maize straw decomposition, improve quality, this study from the low temperature (average temperature of -7 DEG -8 DEG) set for straw returning soil, mining ecological environment in cattle and sheep dung, rotting leaves samples for bacteria the source material, through enrichment culture, carbon source of subculture and the process of cold acclimation, screening of low temperature (4 DEG to 10 DEG) composite microbial system efficient degradation of corn straw, the straw decomposition and clear characteristics, bacteria Composition characteristics, stability and adaptability and application effect, for low temperatures in North China (4 to 10 DEG C) of maize straw fast, high decomposition efficiency and provide a theoretical basis for rational use, and provide strain resources. The main results are as follows: 1. screening of corn straw degrading composite microbial system: low temperature using enrichment culture. Of subculture, low temperature acclimation by generation techniques on the 128 samples collected from corn straw soil, alpine cattle and sheep dung, rotting leaves such as bacteria source materials were selected, 7 low temperature (10 DEG C), temperature (15 DEG C), two type strains and 20 low temperature (10 DEG C) high efficient strains. Further through the analysis of the degradation rate of corn straw and cellulase activity of different strains of dynamic algebra, finally obtain a set at a low temperature (4 DEG to 10 DEG) under High Cellulase Producing Strain GF-, composite and stable degradation of corn straw 20, in the 12h after inoculation into the logarithmic growth phase, cellulase, corn stalk decomposition into simple hydrocarbons or esters. Cultured 15d at 10 DEG C under low temperature conditions, corn straw degradation rate reached 32.29%, compared with a composite microbial system No.8 (the research group in 2013 selected 15 Deg. C degradation rate was 22.02%) 10.27%.2. high efficient low temperature degradation of corn straw composite strains of species composition diversity: a composite microbial system GF-20 consists of 22 kinds of bacteria and 4 fungi. Bacteria were Azonexus hydrophilus, Azospira oryzae, Variovorax boronicumulans, Hydrogenophaga caeni, Herbaspirllum frisingense, Arcobacter cloacae, Pseudomonas fuscovaginae, Cellvibrio Mixtus subsp.mixtus, Seleniivibrio woodruffii, Rhodococcus qingshengii Terribacillus, saccarophilus, Planococcus antarcticus, Bacillus licheniformis, Bacillus tequile NSIS, Clostridium populeti, Clostridium xylanolyticum, Paludibacter propionicigenes, Pedobacter agri, Sphingobacterium cladoniae and Uncultured bacterium. Lotharella globosa Trichosporon is the main fungi, sp., Trichosporon loubieri and Chlamydomonas leiostraca.3. of corn straw degrading composite microbial system of low temperature stability and adaptability: a composite microbial system GF-20 in corn straw decomposition process in central changes is relatively stable, there is the key strain stability; at different temperature (4 DEG -30 DEG) and pH value (6.0-9.0) under the condition of subculture, the temperature (0 DEG -40 DEG) under continuous cultivation, the composite microbial system properties, compositions and functions are stable, has strong stability and adaptability; carbon source for corn straw, nitrogen source when ammonium sulphate composite strains GF-20 strain which is more stable, induced by complex strains producing cellulase, promote the decomposition of substrate Critical dilution; gradient composite microbial system GF-20 was 1014 times higher than that of.4. corn straw degrading composite microbial system in low temperature application: 10% inoculation amount, pH value 6.5, corn straw and ammonium sulfate added than 40:1, 10 degrees Celsius temperature, osmotic concentration of 2% under the conditions of fermentation decomposition agent GF-20 can promote the decomposition of good results at 4 C, 10 C and 30 60d solution rot C conditions, corn straw degradation rate were 29%, 36.75% and 59.47%, 7.21% higher than the control lvkang agents, 9.78% and 10.26%; available nitrogen, available phosphorus, available potassium and organic matter content were increased by 0%-5.32%, 5.26%-14.17%, 2.07%-3.57% and 0.44% ~ 2.94%; significantly increased soil catalase, urease, alkaline phosphatase, invertase, B- glucoside enzyme and cellulase activity increased; soil microbial richness index and diversity index, the new Candidatus saccharibacteria and Planctomycetia. Material group, Herbaspirillum Bacillus sp., promoting lusitanum, fibrivorans and Cellvibrio increase the functional microbes such as albomyces and Melanocarpus, agent GF-20 on maize seed germination rate had no effect. The corn straw degrading composite microbial system GF-20 with low temperature stable corn straw decomposition activity and bacterial composition, in the condition of low temperature (4 DEG to 10 DEG C) under effectively accelerate straw decomposition process, improve the nutrient content and enzyme activity of soil available, increase soil microbial diversity, with good development potential and application prospect.

【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S141.4

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